A platform to discuss the methods and technologies involved in DNA sequencing. | Contact experts in DNA Sequencing to get answers
1,316 views 471 posts
Questions related to DNA Sequencing
Seeking to learn the steps for using the BLAST tool in NCBI for designing primers for make a PCR a specific SNP.
06 June 2014 9,398 5 View
I made a construct of unknown gene in a vector and sequenced. The sequenced data did not match with the expected sequence. How can I troubleshoot this?
06 June 2014 6,799 2 View
Has anyone used the Genewiz service for DNA synthesis? I don´t have any references and I would like to know your experiences.
06 June 2014 842 4 View
I'm looking for a method to improve the yield of a p15A plasmid. I seem to recall hearing about the use of antibiotics to block protein expression or cell division, while still allowing plasmid...
06 June 2014 2,805 5 View
I need to determine if there are any sites for the enzyme AscI in this vector for which I need the entire sequence. However, it does not seem to be available anywhere.
06 June 2014 9,760 5 View
I need to transform E. coli TAP56 with a plasmid of 5.6 kbp. The CaCl2 transformation protocol was working fine earlier. But for last few weeks this method is not working at all. We have sequenced...
06 June 2014 9,268 5 View
I'm trying to replicate the results of the DNA labeling and single molecule imaging section but have not had luck getting the same results as the publication. I was wondering if someone who has...
06 June 2014 2,989 1 View
I am targeting nuclear genes of vertebrate species. The amplicon size is 300 bp. In my PCRs I get a single band on 2% gel, but in DNA sequencing only the reverse primer generated sequences are...
06 June 2014 5,477 13 View
I am using direct sequencing (with no cloning, not possible) and not sure if it is necessary that I sequence with both the forward and the reverse for each sample. In theory, if I just used the...
06 June 2014 8,303 3 View
03 June 2014 9,340 5 View
Criteria for selection of a gene for phylogeny - let it be the case of HCV ?
05 May 2014 6,611 3 View
We need a 10-mer DNA sequence which won't hybridize with any human mRNA sequence. But I design a few and always can find it 100% match in human mRNA database. Can anyone help me on this?
05 May 2014 9,079 1 View
Microbial prokaryotic identification is usually carried by nucleotide sequencing?
05 May 2014 8,963 3 View
I need to concentrate 500bp frag DNA into 10ul.
05 May 2014 3,857 4 View
We've done lots of sequencing of PCR products and plasmids, but we are trying to perform some genomic sequencing of plant DNA in our lab now. I'm just curious if anyone has any advice for getting...
05 May 2014 3,440 0 View
I am trying to isolate and purify the DNA for Next Gen Sequencing after doing Chromato-immunoprecipitation (ChIP). I have tried doing purification using Qiagen based columns (high loss of DNA) and...
05 May 2014 3,299 21 View
I am student at Nottingham University, working on a project on Yam in Malaysia. I am having difficulty finding where to collect samples, because I am an international student. I would be very...
05 May 2014 8,912 1 View
I am doing chip-seq. But the q-PCR result after establishment of the chip-seq library for the same loci is different from before chip-seq library. Who can tell me the potential reasons and how can...
04 April 2014 5,254 6 View
For my bacteria gene, after I got a sequence from the company I compared with the original gene from the gene bank. The result was that my gene differed by just one nucleotide, so that changed 1...
04 April 2014 7,776 3 View
I am leading a cloning using pBluescript vector. When I arrived at the ligation step I realized that it involves specific working conditions applied in the case where cohesive ends are implicated...
04 April 2014 8,353 7 View
Subtelomeric DNA regions are adjacent to the telomeric regions of chromosomes. These bear a number of epigenetic marks.
04 April 2014 8,595 2 View
DNA-based identification have been used in nematology for decades. If I use several common DNA markers (eg. SSU and LSU D2D3) to identify one group of nematodes, may I call it DNA barcoding?
04 April 2014 1,318 3 View
I was given a PCR product with BamHI sites conveniently located near, but not at, the ends: BamHI BamHI I then cloned the BamHI fragment into pUC19 as follows: First I digested the pUC19 and...
03 March 2014 2,008 5 View
Just wanted to know if any one one had come across multiple missense mutations in a single exon for a single sample. Just got back my Sanger Sequencing results and one of the reads has like 3...
03 March 2014 3,247 7 View